PertaWka 9(1), 119 -123 (1986)

COMMUNICATION I
Determination of Sugars in Soft Drinks by
High Performance Liquid Chromatography
Key words: HPLC, soluble sugars; soft drinks.

ABSTRAK

Kandungan fruktosa, glukosa dan sukrosa dalam minimum ringan terpilih yang terdapat di
Malaysia ditentukan melalui kromatografi cecair prestasi tinggi (HPLC). Minuman ringan yang diuji
mengandungi 8.5 -15.3 g 100 ml- 1 gula larut. Kandungan fruktosa, glukosa dan sukrosa masing-
masing didapati dalam julat 0 - 6. 7, 0 - 6.9 dan 0 - 10.5 g 100 ml -1. Didapati bahawa pada minu-
mam ringan yang spesifik, perbezaan adalah lebih besar antara kandungannya untuk gula in-
dividu daripada kandungannya bagijumlah gula.

ABSTRACT

The fructose, glucose and sucrose contents of selected soft' drinks available in Malaysian
markets were determined by high performance liquid chromatography (HPLC). The soft drinks tested
had a soluble sugar content of between 8.5 to 15.3 g 100 ml- 1• The average fructose, glucose and
sucrose contents were found to be in the ranges of 0 - 6. 7, 0 - 6.9 and 0 - 10.5 g 100 ml I respectively.
The content of individual sugars were found to be more variable than the content of total sugar in dzj-
ferent samples of a specific soft drink.
INTRODUCTION MATERIALS AND METHODS

The past few years has seen the mushroom-

Materials
ing of the soft drink industry in Malaysia.
Besides the well known soft drinks like Seven Up Samples of soft drinks were bought from
and Coca-cola, many new varieties which are local supermarkets. 4 samples of each drink were
lesser known in-the West, have gained popularity analysed. The drinks selected were: (i) OI;ange
in the local market. These include chrysan- crush (F & N), (ii) Coca-cola, (iii) A & W root
thenum and herbal tea, longan-winter melon beer, (iv) Dads root beer, (v) Schweppes orange,
and sugar cane drinks, as well as fruit juices like (vi) Fanta (Gedep Merk) vruchtenlimonade si-
guava, mango and mango-pineapple juice. naasappel, (vii) Yeo's longan-winter melon
drink, (viii) Drinho chrysanthenum tea (ix)
Interest in the sweetened soft drinks, both Drinho sugar cane drink, (x) Joy mango juice (xi)
carbonated and non-carbonated, arises from two Joy guava juice, (xii) Delite mango juice (xiii)
considerations. Firstly, the analysis of the sugar Dewi mango juice, (xiv) Sunjus orange juice (xv)
contents of locally produced soft drinks by high Green Spot orange juice drink and (xvi) Drinho
pressure liquid chromatography (HPLC) has not herbal tea.
been undertaken before. Secondly, from a nutri-
tional view point, sucrose intake at high levels Analyses of Sugars
has been implicated in such health problems as
diabetes mellitus (Cohen et al., 1974), athero- A HP 1048B liquid chromatograph with an
sclerotic heart disease (Kaufmann et al., 1967), RI detector was used. The method was based on
carbohydrate malabsorption syndromes (Do- that of Hurst et al., 1977. The column was an
naldson and Grybosky, 1973) and dental caries NH 2 polar bonded phase column, 10 J1 m (250
(Hartles, 1967). mm X 4.6 mm I.D.). The mobile phase was
 M.A. AUGUSTIN AND K.L. KHOR

acetonitrile:water (85:15) and the flow rate was SUGAR STANDARD JOY GUAVA JUICE
2.5 em min -1. The injection volume was 10 J.L 1.
Identification and quantification of sugars were ..
.,.,
done by comparing retention times and peak r 00

areas of samples to peak areas of standards as '"

00
CN

CN
peak area was directly proportional to the con-
centration of the standard throughout the con- G G

centration range used.

""o
Soft drinks were analysed as received with '"
minor sample preparation. Carbonated drinks
were decarbonated by vigorous agitation of solu-
tions with a glass rod for 5 min. For the analysis,
2 ml of the drink was diluted to 10 ml in a
volumetric flask. The diluted drink solution was
s
filtered through C-18 Sep Pak cartridges and
""
0
.,.;
0.45 J.l m filters prior to injection into the HPLC. .,., '"0f-<
CN
0
""' 0....
""6 '"
Recovery studies were carried out by spiking soft ~
l@ 10~
rr
'"
drinks with known amounts of a standard sugar
solution containing glucose, fructose and
sucrose. For spiking experiments, 2 ml of stan-
dard sugar solution containing 1.6 X 10 -2 g Fig. 1: HPLC chromatograms of (I) standard
ml -I of each of the sugars glucose, fructose and sugar solution and (ill diluted Joy guava
sucrose was mixed with 1 ml of the undiluted juice. (F - fructose; G - Glucose;
drink and the mixture was made up to volume in S - sucrose)
a 10 ml volumetric flask. These studies showed
that the recovery for fructose, glucose and drinks analysed by Martin-Villa et ai. (1981).
sucrose for the range of soft drinks was between Thus, it is evident that the range of total sugars
91 to 107 %. However, the variation in the % in soft drinks found in locally available soft
recovery between individual sugars in a specific drinks is in a similar range to that reported.
drink was much less and averaged about 2.6%. There are however more marked differences in
the contents of individual sugars.
RESULTS AND DISCUSSION
There is a wide variation in the content of
Sugar Content of Soft Drinks individual sugars present in the range of soft
drinks analysed. The average fructose and
Fig. 1 shows two chromatograms obtained glucose contents were found to be 0 - 6.7 and
using the HPLC conditions described. Table 1 0- 6.9 g 100 ml I respectively. The average
shows the data obtained for the soft drinks sucrose content was in the range of 0 - 10.5 g 100
analysed. The total sugar content was found to ml - 1 • One of the main reasons for the differences
be between 8.5 to 15.3 g 100 ml -I. The lowest in the content of individual sugars is the dif-
sugar content was found in Drinho sugar cane ferent amounts of corn sweeteners and sucrose
drink while .the highest was found in one batch of used in the manufacture of different drinks.
Joy guava juice. The average value for total Labels on the soft drink containers indicated
sugar in the drinks analysed in this work was that in some drinks corn sweeteners (which in-
found to be 12 g 100 ml -I. Vidal-Valverde et ai. cludes high fructose-glucose syrups) or/ and
(1985) found that the total amount of sugars sugar was used in the manufacture of the drinks.
ranged from 9.2 to 14 g 100 ml - 1 in soft drinks The labels on some of the other drinks stated
and fruit nectars he analysed. A total sugar con- only that 'sugar' was used as an ingredient. The
tent of 10 to 12 g 100 ml -I was found in most definitions of sugar and sweeteners for soft

120 PERTANIKA VOL. 9 NO. I. 1986

 SUGARS IN SOIT DRINKS

TABLE 1
Soluble sugars in soft drinks

Fructose Glucose Sucrose Total Sugars

Type of Drink Container
g 100 ml- I g 100 ml- I g 100 ml- I g 100 ml- I

Orange Crush (F&N) Bottle 6.57 ± 0.09 6.31 ±0.23 < 0.46 13.00±0.41
Coca-Cola Bottle 5.65±0.57 5.57 ± 0.05 <0.16 11.35±0.12
Can 3.40 ± 0.25 3.65 ± 0.25 3.60 ± 0.09 10.65 ± 0.43
A & W root beer Can 6.62 ±0.12 4.71±0.11 n.d. 11.32 ± 0.23
Dads root beer Can 3.75±0.10 3.74 ± 0.09 4.25 ± 0.52 11.74 ± 0.35
Schweppes orange Can 6.58 ± 0.28 6.36 ±0.24 < 1.19 13.41 ± 0.38
Fanta (Gedep Merk) Can 6.69 ± 0.44 6.50 ± 0.41 <0.91 13.67 ± 0.92
vruchtenlimonade
sinaasappel
Yeo's longan-winter Tetra-pak 0.43 ± 0.02 0.42 ± 0.02 9.59 ± 0.46 10.45 ± 0.47
melon drink
Drinho chrysanthenum tea Tetra-pak <0.06 <0.05 10.48 ± 0.50 10.53 ± 0.50
Drinho sugar cane drink Tetra-pak (a) 1.41 ± 0.01 1.97 ±0.01 5.10 ± 0.01 8.47 ± 0.01
(b) 0.58 ± 0.09 0.55 ± 0.12 9.19±0.18 10.20 ± 0.35
Joy mango juice Tetra-pak 3.53±0.13 3.11 ±0.08 5.64 ± 0.14 12.28 ± 0.32
Dewi mango juice Tetra-pak (a) 6.23 ± 0.15 6.28 ± 0.44 2.42 ± 0.34 14.93±0.93
(b) 5.60±0.17 5.01 ± 0.11 2.10±0.87 12.72±0.32
Delite mango-pineapple Tetra-pak 6.04 ± 0.16 6.06 ± 0.23 < 1.47 12.64±0.16
drink
Sunjus orange juice Tetra·pak 6.62 ± 0.11 6.48 ± 0.08 <1.22 14.05 ± 0.16
Joy guava juice Tetra·pak (a) 4.46 ± 0.35 5.27 ± 0.26 5.58 ± 0.56 15.31 ± 0.05
(b) 6.39 ± 0.10 6.16±0.66 0.31 ± 0.02 12.82 ± 0.11
Green Spot orange Tetra·pak (a) 6.41 ± 0.18 6.93 ± 0.46 0.82 ± 0.25 14.44±0.98
juice drink (b) 6.22 ± 0.09 6.08 ± 0.20 <0.57 12.67 ± 0.07
Drinho herbal tea Tetra-pak <0.03 <0.03 9.81 ± 0.20 9.83 ± 0.18

The symbols (a) and (b) refer to analyses of different batches of drinks which were bought at different times. In
drinks, other than those with symbols (a) and (b), the differences in sugar contents between batches were small
and less marked. n.d. = not detectable.

drinks vary from country to country. In the 1964 sugar can also arise because of other factors. In
Soft Drink Regulation (UK), sugar is defined as fruit juice drinks where the juice, concentrate or
'any soluble carbohydrate sweetening matter' puree of the fruit is added as an ingredient, dif-
while the sugar product regulations of the EEC ferences in the contents of individual sugars in
identifies the term sugar with 'sucrose' (Tilley, the drink may be due to inherent differences in
1978). In the Food Regulations 1985 of Malaysia the contents of individual sugars in different
sugar is defined as the food chemically known as fruits. Another contributory factor is the inver-
sucrose and includes granulated sugar, loaf sion of sucrose during storage. It has been stated
sugar, castor sugar and powdered sugar. Sugar that it is quite reasonable to assume that sucrose
should contain not less than 99.5 % of sucrose. undergoes hydrolysis in the acidic pH of soft
drink media (Martin-Villa et al., 1981; Vidal
The differences in the content of individual Valverde et at., 1985).
PERTANIKA VOL. 9 NO. I, 1986 121
 M.A. AUGUSTIN AND K.L. KHOR

In this work, there were also marked dif- ACKNOWLEDGEMENT

ferences in the individual sugar content in dif-
ferent samples of Coca-cola, Drinho sugar cane The authors would like to thank Halim
drink, Dewi mango juice, Joy guava juice and Abdul Rahman and Azman Abu Yamin for their
Green Spot orange juice drink. The pH of these technical assistance in the analysis and The New
drinks was found to be between 2.5 to 4.8. It Straits Time Press (Malaysia) Bhd. for provision
would appear that these differences may be part- of funds for the analysis.
ly due to inversion of sucrose during storage. In
the paper by Vidal-Valverde et al., (1985) it was M.A. Augustin and
also stated that a low sucro~e content was pro- K.L. Khor'
bably an indication of long storage time. A
Department ofFood Science,
number of reports have shown that there can be
Faculty ofFood Science and Technology,
significant differences in the individual sugar Universiti Pertanian Malaysia,
content of the same type of drink. Pinalla (1968)
43400 Serdang, Selangor, Malaysia.
found 9.5% sucrose in cola drinks while South-
gate et al., (1978) found 0.5% sucrose in cola
drinks. In recent analysis, the sucrose level in
REFERENCES
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"Department of Human Development Studies. Faculty of Agriculture.

122 PERTANIKA VOL. 9 NO.1, 1986

 SUGARS IN SOIT DRINKS

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PERTA IKA VOL. 9 NO. I, 1986 123